Effects Of Advanced Glycation End Products On Oxidative Stress And Apoptosis In Cultured Bovine Trabecular Meshwork Cells

ObjectiveTo investigate the effect of advanced glycation end products (AGEs) on apoptosis and proliferation of cultrued bovine trabecular meshworks cells (TM cells) and to further explore the relationship between AGEs and Primary open angle glaucoma (POAG), probe the pathogenes of glaucoma induced by diabetic.MethodsCultured bovine TM cells were identified by their origin (expression of NSE.Ⅷ-factor and FN). The third generation of cells were selected, and different concentrations (0、50、100、200μ g/mL) AGEs was added into the medium for 96 hours, or treated with AGEs (200 μg/mL) for different times (48,72,96h). MTT [3-(4,5-Dimethylthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT] was performed to detect the cell proliferation;Flow cytometry (FCM) was performed to detect the cells apoptosis; the level of ROS was evaluated by the 2’,7’-dichlorofluorescein diacetate (DCFH-DA) method.ResultsTake the third generation bovine TM cells for characterization. FN-stained and NSE-stained by immunofluorescence was positive. Ⅷ-facter stained by immunofluorescence was negative. The second generation bovine TM cells get into logarithmic growth phase on the third day and maintain about three days. The method of MTT display the AGEs may inhibit the proliferation of bovine TM cells in vitro in a dose-dependent manner (P<0.05). Flow cytometry showed that the AGEs induced cultrued bovine TM cells apoptosis in a time and concentration-dependent manner, at the same time AGEs significantly increased the production of reactive oxygen species in a dose-dependent manner (P<0.05).ConclusionThe third generation bovine TM cells in vitro should be chosed into experiments of glaucoma AGEs can inhibit the proliferation of bovine TM cells in vitro; AGEs can caused an increase in levels of ROS in bovine TM cells in vitro, which may be involved in apoptosis of bovine TM cells in vitro induced by AGEs.